CN108929672A - It is a kind of using shrimp shell as carbon quantum dot of carbon source and preparation method thereof and detection ascorbic acid in application - Google Patents

It is a kind of using shrimp shell as carbon quantum dot of carbon source and preparation method thereof and detection ascorbic acid in application Download PDF

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CN108929672A
CN108929672A CN201810534230.4A CN201810534230A CN108929672A CN 108929672 A CN108929672 A CN 108929672A CN 201810534230 A CN201810534230 A CN 201810534230A CN 108929672 A CN108929672 A CN 108929672A
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quantum dot
preparation
ascorbic acid
carbon quantum
fluorescence
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CN108929672B (en
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台德艳
王丽珍
刘金水
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Anhui Normal University
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Anhui Normal University
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    • C09KMATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y20/00Nanooptics, e.g. quantum optics or photonic crystals
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B82NANOTECHNOLOGY
    • B82YSPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00Manufacture or treatment of nanostructures
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6432Quenching

Abstract

The invention discloses a kind of using shrimp shell as carbon quantum dot of carbon source and preparation method thereof and the application in detection ascorbic acid.It will be distributed in ultrapure water after ash content grinding of the shrimp shell after high temperature sintering, and obtain carbon quantum dot solution after centrifugation, filtering, dialysis.Utilize carbon quantum dot and Cr6+Composite fluorescence probe is constructed, constructs linear relationship curve using fluorescence analysis, and then realize the quantitative detection of Ascorbic Acid.This method is low in cost, high sensitivity, linear relationship is good, easy to operation, selectivity is preferable.

Description

It is a kind of that Vitamin C as carbon quantum dot of carbon source and preparation method thereof and is being detected using shrimp shell Application in acid
Technical field
The present invention relates to a kind of using shrimp shell as carbon quantum dot of carbon source and preparation method thereof and in detection ascorbic acid Using.
Background technique
Carbon quantum dot (Carbon Quantum Dots, abbreviation CQDs or CDs) be it is a kind of have to semiconductor-quantum-point it is similar The environmentally friendly fluorescent nano material of optical property.Despite the up-and-coming youngster in nanometer large family, but it is because it has Excitation spectrum is continuous and broad, unitary excitation, and polynary transmitting, fluorescence is strong and the excellent fluorescence properties such as stability is good, and goes back With partial size small (diameter be less than 10nm), molecular weight is low, bio-toxicity is low, preparation cost is cheap, is easy to functional modification and anti- Answer the advantages such as mild condition, just gradually replacing metal quantum point and organic dyestuff be widely used in photoelectric device, heavy metal from The fields such as sub- detection, chemical sensor, photocatalysis, biomarker and cell imaging.
L-AA (Ascorbic Acid, abbreviation AA) is usually called vitamin C (Vitamin C, abbreviation VC), is One of important water soluble vitamin, main function is the normal physiological function for maintaining body, but body itself cannot synthesize, and It can only and must be absorbed from food and drug.Vitamin C is usually had in the complicated metabolic processes such as body oxidation, reduction Acid participates in extensively, and it can promote the growth and formation of antibody, enhances the resistance to various diseases, while being also equipped with A degree of detoxication.Research finds that body lacks ascorbic acid and may result in a variety of strange diseases, the height of content Low degree would generally be as one of trophic analysis and the important indicator of certain medicals diagnosis on disease.
Therefore, it is very heavy for developing the simple and easy ascorbic acid analytic approach of one kind come the detection for life daily bread It wants.Currently, there are many measuring method of ascorbic acid, it has been reported that focusing primarily upon following several method in lot of documents:Electricity Chemical method, titration, high performance liquid chromatography, photometry, enzyme process, fluorescent spectrometry, voltammetry etc..Wherein, certain methods are wanted The experiment condition asked is more harsh and operating technology is higher, and some method and step complexity are cumbersome, and be unfavorable for quickly analyzing wants It asks.
Summary of the invention
The present invention provides a kind of using shrimp shell as carbon quantum dot of carbon source and preparation method thereof and in detection ascorbic acid Application.Utilize carbon quantum dot and Cr6+Composite fluorescence probe is constructed, constructs linear relationship curve using fluorescence analysis, in turn Realize the quantitative detection of Ascorbic Acid.This method is low in cost, high sensitivity, linear relationship are good, easy to operation, selection Property is preferable.
The technical scheme adopted by the invention is as follows:
It is a kind of using shrimp shell as the preparation method of the carbon quantum dot of carbon source, include the following steps:It is dried after shrimp shell is cleaned, so 4.5~6h of high-temperature calcination obtains ash content at 220~240 DEG C afterwards;Ultrapure water is added to by ash content grind into powder, and by powder Middle ultrasonic disperse, and carbon quantum dot solution is obtained after centrifugation, supernatant liquid filtering, filtrate dialysis.
Further, the temperature of the drying is 80 DEG C;The temperature and time of the high-temperature calcination is respectively preferably 230 ℃、5h。
The ratio of the powder and ultrapure water is 2g:80~120mL;Preferably 2g:100mL.
The time of the ultrasound is 0.8~1.5h;Preferably 1h.
The condition of the centrifugation is:9000~10000rpm/min of centrifuge speed, centrifugation time are 25~40min;Into One step, preferably:Centrifuge speed 9500rpm/min, centrifugation time 30min.
The filtering refers to that by 0.22 μm of micro-pore-film filtration, what the dialysis referred to is 3500Da's through molecular cut off 10~15h of dialysis is carried out in bag filter;Preferably dialyse 12h.
The present invention also provides the carbon quantum dots being prepared according to above-mentioned preparation method, are evenly distributed, partial size 3~ 8nm。
For the present invention using cheap shrimp shell as carbon source, gained carbon quantum dot has good fluorescence property and photostability, When excitation wavelength is 340nm, there is strongest fluorescence intensity at 400nm wavelength, and the peak shape of fluorescence emission peak is good.And Preparation process is easy to operate, and used solvent only has water, is a kind of preparation method of green non-pollution.
The present invention also provides the carbon quantum dots being prepared according to the preparation method in detection ascorbic acid Using.
The present invention also provides a kind of detection methods of ascorbic acid, include the following steps:By carbon quantum dot solution and Cr6+ Aqueous solution is mixed to get CQDs/Cr6+Composite fluorescence probe solution, and adjusting pH is 7.0, then to CQDs/Cr6+Composite fluorescence The aqueous ascorbic acid of different final concentrations is added in probe solution, it is strong to test fluorescence of each system under 340nm excitation wavelength Degree;Using the ascorbic acid concentrations within the scope of 25~95 μM as abscissa, the maximum value of the fluorescence intensity at 400-420nm is vertical sits Mark building linearity curve, and then measure the concentration of ascorbic acid in prepare liquid.
Into CQDs plus Cr6+Ion, with Cr6+Ion concentration increases, fluorescence emission spectrum gradually red shift, from 400nm 420nm is moved on to, as shown in Figure 5;As past CQDs/Cr6+Gradually add AA in composite fluorescence probe solution, since AA is Cr in system6+ Ion reduction is to Cr3+Ion leads to fluorescence emission spectrum gradually blue shift, moves on to 400nm from 420nm, as shown in figure 3, to extensive The 400nm emission spectrum of CQDs itself is arrived again.Therefore the maximum value of the fluorescence intensity at 400-420nm is chosen as ordinate building Linearity curve.
Further, the CQDs/Cr6+In composite fluorescence probe solution, Cr6+Final concentration of 140 μM of aqueous solution. The fluorescence intensity of CODs is with Cr6+The increase of concentration, fluorescent quenching degree increase, and work as Cr6+When concentration reaches 140 μM, CODs's Fluorescent quenching reaches 60%;This patent selects Cr6+Concentration reaches 140 μM for detecting AA.Although with Cr6+The continuation of concentration Increase, the fluorescence of CQDs further quenches, but quenching amplitude is little, if Cr6+Concentration is too big, it will more AA are consumed, thus Cause detection AA insensitive.
Further, the linear equation of the linearity curve is Y=637.15+4.75C, and wherein Y is at 400-420nm The maximum value of fluorescence intensity;C is ascorbic acid concentrations, and unit is μM;Linearly dependent coefficient is R=0.993, and detection limit is minimum can To reach 1.14 μM.
Carbon quantum dot provided by the invention is in the application of detection ascorbic acid and the detection method of ascorbic acid, carbon quantum Point and Cr6+In conjunction with rear, the fluorescence intensity of carbon quantum dot is quenched, after adding AA, Cr6+It is aoxidized between the AA of addition Reduction reaction generates Cr3+The fluorescence of ion, carbon quantum dot restores therewith, and the concentration of AA is bigger, and the degree that fluorescence restores is higher, Fluorescence intensity and within the scope of 0~178 μM, at the maximum value of fluorescence intensity of the concentration and system of AA at 400-420nm It is worth linear correlation, linearly dependent coefficient 0.993.The quantitative detection to AA concentration to be measured can be realized in turn.
Carbon quantum dot disclosed by the invention realizes that the principle of AA detection is:Existed using the carbon quantum dot that shrimp shell is prepared as carbon source Excitation map (there is maximum fluorescence intensity at 340nm wavelength), the hair under 340nm excitation wavelength under 400nm launch wavelength Penetrate map (there is maximum fluorescence intensity at 400nm wavelength) and Cr6+The ultraviolet absorpting spectrum generation of ion is significantly overlapped, So as to which fluorescence inner filtering effect (IEF) effectively occurs, so the fluorescence of carbon quantum dot can be by Cr6+Rather than Cr3+Ion Quickly and efficiently quench.And after addition AA, Cr6+Redox reaction occurs between ion and the AA of addition and generates Cr3+From Son, thus the fluorescence of carbon quantum dot can restore.
It is compared with prior art, disclosed by the invention simple using shrimp shell as the environmental protection of the preparation method of the carbon quantum dot of carbon source, Carbon quantum dot and Cr can directly be utilized6+The composite fluorescence probe of ion building realizes the quantitative detection to AA, detection method spirit Sensitivity is high, linear relationship is good, easy to operation, the good, strong antijamming capability of selectivity.
Detailed description of the invention
Fig. 1 is the TEM figure of carbon quantum dot in embodiment 1;
Fig. 2 is the fluorescence emission spectrogram of compound of the carbon quantum dot in the embodiment 1 under different excitation wavelengths;
Fig. 3 is to CQDs/Cr6+The fluorescence emission spectrum after various concentration AA solution is added in composite fluorescence probe solution Figure;
Fig. 4 is the fluorescence intensity level at the maximum value of the fluorescence intensity with AA concentration to detection architecture at 400-420nm The linear relationship chart of building;
Fig. 5 is the Cr that various concentration is added into CQDs6+Fluorescence emission spectrogram of compound afterwards;
Fig. 6 is excitation spectrum, emission spectrum and the Cr of carbon quantum dot6+The uv absorption spectra of ion;
Fig. 7 is Cr6+Ion and the ultraviolet spectrogram being added after AA.
Fig. 8 is CQDs/Cr6+The selectivity and interference--free experiments figure of system detection ascorbic acid.
Specific embodiment
Embodiment 1
It is a kind of using shrimp shell as the preparation method of the carbon quantum dot of carbon source, include the following steps:
It places in an oven, is taken out after drying moisture under 80 DEG C of environment, to naturally cold after weighing the dry shrimp wash clean of 10.0g But it to room temperature, then puts it into crucible and is placed in Muffle furnace the high-temperature calcination 5h at 230 DEG C, it is natural to calcined ash content It is cooled to room temperature rear grind into powder.
Weigh 2.0g powder be added in 100mL ultrapure water be mixed shake up, be put into ultrasound in Ultrasound Instrument make its uniformly Dispersion.Will solution be ultrasonically treated 1h after using supercentrifuge setting revolving speed be 9500rpm/min, centrifugation time 30min, will It is centrifuged and removes large particulate matter, collects supernatant.
By 0.22 μm of micro-pore-film filtration to obtain faint yellow filtering solution, final product solution is using gained supernatant Preceding in molecular cut off is further to dialyse 12h in 3500Da bag filter, takes the solution outside bag filter up to carbon quantum dot (CQDs) solution.
The pattern of CQDs is analyzed using transmission electron microscope picture (TEM), as shown in Figure 1, it can be seen that prepared CQDs is spherical shape, and partial size is mainly distributed in the range of 3~5nm, average in 4nm or so, and is distributed more uniform.
Similar with most of fluorescent carbon quantum dots, CQDs prepared by the present invention also has excitation dependence Fluorescence behaviour Property, as shown in Fig. 2, CQDs emission peak positions and fluorescence intensity be not all as the change of excitation wavelength is from 300 to 360nm Disconnected variation.When excitation wavelength is 340nm, the fluorescence spectrum of CQDs has strongest fluorescence emission peak, peak at wavelength 400nm Shape is good, thus below to AA detect when fluoremetry when excitation wavelength set λex=340nm.This excitation dependence fluorescence row For the surface defect generation for being the optics selection due to various sizes of CQDs with CQDs.
Embodiment 2
Application of the carbon quantum dot solution that embodiment 1 obtains in detection ascorbic acid.
The method of the detection is:The Cr of carbon quantum dot solution and 0.1mL that the embodiment 1 of 1mL is obtained6+Aqueous solution is mixed Conjunction obtains CQDs/Cr6+Composite fluorescence probe solution, Cr6+Final concentration of 140 μM;And 0.6mL PBS buffer solution tune is added Saving pH is 7.0, then to CQDs/Cr6+The aqueous ascorbic acid of different final concentrations is added in composite fluorescence probe solution, After 15min, fluorescence intensity of each system under 340nm excitation wavelength is tested, as shown in Figure 3.With anti-within the scope of 0~178 μM Bad hematic acid concentration is abscissa, and the fluorescence intensity level at the maximum value of the fluorescence intensity at 400-420nm is that ordinate constructs line Linearity curve, as shown in figure 4, an available good linear relationship, linear equation Y=within the scope of 25~95 μM 637.15+4.75C, wherein Y is the maximum value of the fluorescence intensity at 400-420nm;C is ascorbic acid concentrations, and unit is μM;Line Property related coefficient be R=0.993, detection limit minimum can achieve 1.14 μM;And then measure the concentration of ascorbic acid in prepare liquid.
Embodiment 3
CQDs/Cr6+Cr in composite fluorescence probe solution6+The selection of concentration
CQDs prepared by 1.0mL embodiment 1 is taken respectively, is then separately added into the Cr of different final concentrations thereto6+Solution, And 0.6ml PBS buffer solution is added, pH to 7.0 is adjusted, CQDs/Cr is made6+Probe aqueous solution measures above-mentioned body after 10min The fluorescence intensity of system, such as Fig. 5, from fig. 5, it can be seen that having at 300nm using CQDs solution prepared by dry shrimp carbon source very strong Fluorescence, once various concentration Cr is added into the solution6+Later, the fluorescence of CQDs will be quenched sharply, and with Cr6+Concentration Increase, fluorescence intensity gradually decreases, and works as Cr6+Ion concentration reaches 140 μM, and the fluorescent quenching of system reaches 60%.Work as Cr6+ Concentration is more than 140 μM, and the fluorescence intensity of system continues to quench, but it is little to quench amplitude, if with the Cr of higher concentration6+Ion with The compound building fluorescence probe of CQDs, then after adding AA, it will more AA are consumed, it is insensitive so as to cause detection AA, therefore this 140 μM of final concentration of Cr is chosen in invention6+Solution and CQDs are mixed to get CQDs/Cr6+Composite fluorescence probe solution is to detect AA。
Embodiment 4
Carbon quantum dot inquires into the detection mechanism of AA
For the mechanism that research system fluorescence gos up, the present invention continues to have studied relevant fluorescence, uv-spectrogram.From Fig. 6 In as can be seen that carbon quantum dot under 400nm launch wavelength excitation map (at 340nm wavelength have maximum fluorescence intensity), In the transmitting map (there is maximum fluorescence intensity at 400nm wavelength) and Cr under 340nm excitation wavelength6+The UV absorption figure of ion Spectrum is generated and is significantly overlapped, so as to which fluorescence inner filtering effect (IEF) effectively occurs, so the fluorescence of carbon quantum dot can By Cr6+Ion rather than Cr3+Ion quickly and efficiently quenches.And after addition AA, Cr6+It is sent out between ion and the AA of addition Raw redox reaction generates Cr3+Ion, thus the fluorescence of carbon quantum dot can restore.
From Fig. 7 it can also be seen that hexavalent chromium has wider ultraviolet characteristic absorption peak at 270nm and 372nm respectively, When 12.5 μM of AA are added, the ultraviolet peak absorbance at 372nm can decline, and illustrate that AA has reduced the Cr of a part at this time6+ Ion, so that ultraviolet peak absorbance can weaken;When 50 μM of AA are added, the ultraviolet peak at 372nm, which completely disappears, to be loseed, explanation Cr6+Redox reaction and Cr has occurred in ion and enough AA6+Ion has almost been reduced into Cr3+Ion, to press down Cr is made6+Ion pair carbon quantum dot Quenching of fluorescence, and carbon quantum dot fluorescence is restored.
Embodiment 5
Selectivity experiment
One is stablized excellent fluorescence probe, it is necessary to have preferable selectivity and anti-interference ability.It is such glimmering in order to probe into The anti-interference ability and selectivity of light nano material, the present invention have selected some Common materials gallic acids (GA), L- tryptophane (L-Try), bovine serum albumin(BSA) (BSA), sucrose (Suc), maltose (Mal), tryptophan (Trp), urea (Uera), thiocarbamide (Thi), dopamine (DA) does Choice tests.The final concentration of all of above substance and AA are 178 μM, and experimental result is as schemed Shown in 8, as past CQDs/Cr6+When the biomolecule of same concentrations being added in system, AA, which is added, makes the fluorescence rise degree of system most Greatly, and in addition to AA, other ions or biomolecule are to CQDs/Cr6+The rise degree of system fluorescence can almost be ignored.To sum up The experimental results showed that Cr6+Ion is in quenching CQDs fluorescence and AA rise CQDs/Cr6+When the fluorescence of probe, all have well Selectivity and anti-interference.
It is above-mentioned that Vitamin C as carbon quantum dot of carbon source and preparation method thereof and is being detected using shrimp shell to a kind of referring to embodiment The detailed description that application in acid carries out, is illustrative without being restrictive, can enumerate according to limited range several A embodiment, therefore the change and modification in the case where not departing from present general inventive concept, should belong within protection scope of the present invention.

Claims (10)

1. a kind of using shrimp shell as the preparation method of the carbon quantum dot of carbon source, which is characterized in that include the following steps:Shrimp shell is cleaned After dry, then 4.5~6h of high-temperature calcination obtains ash content at 220~240 DEG C;Add by ash content grind into powder, and by powder Enter into ultrapure water ultrasonic disperse, and obtains carbon quantum dot solution after centrifugation, supernatant liquid filtering, filtrate dialysis.
2. preparation method according to claim 1, which is characterized in that the ratio of the powder and ultrapure water is 2g:80~ 120mL。
3. preparation method according to claim 1 or 2, which is characterized in that the time of the ultrasound is 0.8~1.5h.
4. preparation method according to claim 1 or 2, which is characterized in that the condition of the centrifugation is:Centrifuge speed 9000~10000rpm/min, centrifugation time are 25~40min.
5. preparation method according to claim 1 or 2, which is characterized in that the filtering is referred to by 0.22 μm of micropore Film filtering, it is described dialyse refer to through molecular cut off be 3500Da bag filter in carry out 10~15h of dialysis.
6. the carbon quantum dot that preparation method according to claim 1 is prepared.
7. application of the carbon quantum dot that preparation method according to claim 1 is prepared in detection ascorbic acid.
8. a kind of detection method of ascorbic acid, which is characterized in that the carbon that preparation method described in claim 1 is prepared Quantum dot solution and Cr6+Aqueous solution is mixed to get CQDs/Cr6+Composite fluorescence probe solution, and adjust pH be 7.0, then to CQDs/Cr6+The aqueous ascorbic acid of different final concentrations is added in composite fluorescence probe solution, tests each system and swashs in 340nm Send out the fluorescence intensity under wavelength;Using the ascorbic acid concentrations within the scope of 25~95 μM as abscissa, the fluorescence at 400-420nm is strong Fluorescence intensity level at the maximum value of degree is that ordinate constructs linearity curve, and then measures the concentration of ascorbic acid in prepare liquid.
9. according to the method described in claim 8, it is characterized in that, the CQDs/Cr6+In composite fluorescence probe solution, Cr6+Water Final concentration of 140 μM of solution.
10. according to the method described in claim 8, it is characterized in that, the linear equation of the linearity curve is Y=637.15+ 4.75C, wherein Y is the maximum value of the fluorescence intensity at 400~420nm;C is ascorbic acid concentrations, and unit is μM;It is linearly related Coefficient is R=0.993, and detection limit is minimum to can achieve 1.14 μM.
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